人杀菌/渗透增强蛋白N末端片段表达载体的构建

The Construction of Human Bactericidal/Permeability Increasing Protein N-terminal Fragment Expression Vector

目的：建立杀菌／渗透增强蛋白（BPI）N-端193个氨基酸的重组表达质粒。方法：利用RT-PCR的方法从HL-60细胞内扩增出BPI氨基端1～193个氨基酸的基因序列，克隆入T载体。将BPI 193基因片段定向克隆入原核表达载体pET-28a中，构建重组的原核表达质粒pET-BPI 193，转化大肠杆菌BL 21菌株。结果：从HL-60细胞中扩增得到579bp的BPI 193基因片段，构建了T-BPI 193亚克隆和PET-BPI 193重组表达质粒。结论：成功构建了pET-BPI 193重组表达质粒。

Objective To construct a prokaryotic expression vector encoding first 193 amino acids of BPI N-terminal. Methods Total RNA was extracted from HL-60 and then was amplified by using RT-PCR. The PCR product was cloned into pMD18-T vector. BPI 193 eDNA was directly inserted into pET-28a plasmid with T4 DNA ligase, pET-BPI 193 recombinant expression vector was transformed into competent E. coli BL 21 and protein expression was induced by IPTG. Results A 579 bp fragment was amplified by RT-PCR. BPI eDNA was inserted into pET-28a. Conclusions pET-BPI 193 recombinant expression vector was constructed successfully.